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Original article
2021
:14;
202107
doi:
10.1016/j.arabjc.2021.103226

Green method development approach of superheated water liquid chromatography for separation and trace determination of non-steroidal anti-inflammatory compounds in pharmaceutical and water samples and their extraction

,
a Department of Chemistry, Faculty of Science, King Abdul-Aziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia
b Chemistry Department, Faculty of Science, King Abdulaziz University, P.O Box 80200, Jeddah 21589, Saudi Arabia

⁎Corresponding author. laalkhatib@kau.edu.sa (Lateefa A. Al-Khateeb)

Received: , Accepted: ,
© The Author(s) 2021
Disclaimer:
This article was originally published by Elsevier and was migrated to Scientific Scholar after the change of Publisher.

Abstract

Non-steroidal anti-inflammatory drugs (NSAIDs) are pharmaceutical compounds with anti-inflammatory, analgesic, and antipyretic effects. Herein, a simple and rapid high-temperature liquid chromatography and superheated water chromatography method was developed and validated for the trace determination of NSAID residues of ketoprofen, naproxen, sodium diclofenac, and ibuprofen in water samples. The NSAIDs were separated in less than five minutes using buffered distilled water as the mobile phase and ODS Zirconia RP-C18 column as the stationary phase. Linearity was observed in the van’t Hoff plots of the tested drugs by employing a low acetonitrile percentage (20% ACN) in the mobile phase, without any significant changes in their retention mechanisms. However, nonlinear van’t Hoff plots were obtained for the superheated water chromatography data of the tested drugs because of significant changes in their retention factors, transition stage of the stationary phase, or the mobile phase properties. The limits of detection for ketoprofen, naproxen, sodium diclofenac, and ibuprofen were 14, 2, 4.2, and 32 µg L−1, respectively, and their limits of quantification were 44, 8, 12, and 98 µg L−1, respectively. The accuracy and precision parameters were determined for selected drugs, where the relative standard deviations were in the range of ± 0.2179–2.6741%. In addition, these conditions were employed for the removal of NSAIDs from the water samples using carbon nanotubes. The proposed system was applied for the separation and analyses of drugs in water and pharmaceutical samples, and acceptable recoveries of 90.48–98.15% for the water samples and 99.9–100.08% for the pharmaceutical samples were obtained.

Keywords

NSAID
Superheated Water Chromatography (SHWC)
Trace detection
High-Temperature Liquid Chromatography (HPLC)
Carbon nanotube
Water samples
1

1 Introduction

Non-steroidal anti-inflammatory drugs (NSAIDs) are pharmaceutical compounds with anti-inflammatory, analgesic, and antipyretic effects, and are used in treating various human diseases worldwide including reducing the risk of Alzheimer's disease and in veterinary medicine (Bhattacharya et al., 2013, Christensen, 1998, Imbimbo et al., 2010, Jaturapatporn et al., 2012, Kinney et al., 2006, Su & Chen, 2008). However, NSAID overdose or their long-term use can result in upper gastrointestinal (GI) irritation, development of gastric ulcers, dyspepsia, and bleeding, as well as some cardiovascular effects, which are the some of the major side effects of this class of drugs. Of these, the overdose or long-term use of even acute or chronic from of NSAIDs can cause GI irritation (Su & Chen, 2008) and cardiovascular complications (Bhattacharya et al., 2013). Therefore, NSAID residues in biological matrices such as urine, plasma, or blood of the individuals administered prescribed NSAIDs have been monitored (Imbimbo et al., 2010, Jaturapatporn et al., 2012). Additionally, environmental contamination due to the presence of these drugs occurs because NSAIDs are weak acids with pKa values of 3–5. High solubility of NSAIDs in water (Alquadeib, 2019) and poor degradability can cause unintended long-term effects on the ecosystem and humans (Eslami et al., 2015, Tanwar et al., 2015). These compounds are not regulated as hazardous pollutants but are found in trace quantities ranging from nanograms to micrograms per litre owing to the increasing release of NSAIDs into the water environments due to human usage, excretion, or improper disposal. This environmental contamination of drinking water can be hazardous to human health as many anti-inflammatory 2-arylpropionic acid derivatives are persistent pollutants that are resistant to biological degradation and can pose a threat to the aquatic life (Baranowska & Kowalski, 2012, Khetan & Collins, 2007, Nikolaou et al., 2007). Therefore, improved water purification technologies and water treatment processes employing chemicals are urgently required to ensure the purity of drinking water by the efficient removal of water pollutants including pharmaceutical pollutants. Many techniques for water purification that containing pharmaceutical drugs impurities (Feng et al., 2013, World Health Organization, 2012). These methods are limited by lengthy procedures, high expense, and contamination of water sources by different chemicals. Nanoparticles have been extensively employed to remove pharmaceutical compounds, heavy metals, bacteria, and other pathogens present at low concentrations in complex matrices from different water samples (Al-Khateeb et al., 2021, Bagheri et al., 2016, Khoeini Sharifabadi et al., 2014, Mukaratirwa‐Muchanyereyi et al., Paltiel et al., 2016, Sotelo et al., 2012, Tiwari et al., 2008, Aguilar-Arteaga et al., 2010), and various methods have been proposed for the detection of these compounds. The removal of these toxic compounds from wastewater is necessary for various health and environmental applications. Many conventional methods such as reduction, precipitation, adsorption, oxidation, and ion exchange have been used. However, adsorption has been found to be the most suitable method because of its high efficiency and economics (Abdel Ghafar et al., 2015, Gupta et al., 2016, Moradi et al., 2014, Sadegh et al., 2016). Multi-walled carbon nanotubes (MWCNTs) have been successfully used as cartridge solid-phase extraction (SPE) sorbents for the extraction of different analytes such as NSAIDs (Veclani et al., 2020) (Dahane et al., 2013, Liang et al., 2014, Wan Ibrahim et al., 2018).

Several techniques have been used for the preconcentration and clean-up of analytes for the detection and quantification of NSAIDs in different matrices for toxicological and therapeutic applications (Baranowska & Kowalski, 2012, Buchberger, 2011, Eric-Jovanovic et al., 1998, Musumarra et al., 1983, Nasal et al., 1997, Sotelo et al., 2012). High-performance liquid chromatography (HPLC) is the more commonly used method for the analysis of NSAIDs in water, biological samples, and pharmaceutical formulations with various detection (Martın et al., 1999, Bhattacharya et al., 2013, Ascar et al., 2013, Cserháti & Szőgyi, 2012, Fillet et al., 1999, Klimeš et al., 2001, Koba et al., 2010, Kubala et al., 1993, Mulgund et al., 2009). Additionally, HPLC coupled with various methods such as solid-phase microextraction (SPME)-HPLC (de Oliveira et al., 2005), SPE-liquid chromatography (Sarafraz-Yazdi et al., 2012, Wang et al., 2018), thin-film (TF)-SPME with LC-tandem mass spectrometry (LC-MS/MS) (Krummen et al., 2004, Hirai et al., 1997), SPME with polyethylene glycol-grafted MWCNTs using gas chromatography (GC) with flame ionisation detector (de Oliveira et al., 2005), SPE-GC–MS/MS (Costi et al., 2008), hollow-fibre liquid-phase microextraction (HFLPME)-HPLC for antiretroviral drugs in different waters samples (Payán et al., 2009), molecular imprinting solid-phase microextraction with HPLC (Togunde et al., 2012, Payán et al., 2011), electro-membrane extraction using HPLC with diode array (Debska et al., 2005) and fluorescence detection (Farrington & Regan 2007), and potentiometric (Parham et al., 2009), conductometric (Payán et al., 2011), and voltametric methods (Ni & Kokot 2008) have been reported. Although these methods are costly and time-consuming, they offer many advantages such as high sensitivity and identification of the compound. However, LC-MS is expensive and is not available in many laboratories. In addition, the method employing GC–MS requires derivatisation before the analysis of many pharmaceuticals, and thus requires a lengthy sample preparation time.

Numerous studies have investigated the effects of varying the organic phase percentage in the HPLC mobile phase on the retention and selectivity of compounds (Al-Khateeb & Smith, 2009, Al-Khateeb, 2021, Leyla et al., 2019, Al-Khateeb and Hakami, 2020). However, a decrease in the percentage of the organic solvent in the mobile phase and increase column temperature reduces the analysis time, making it more demanding iif the stationary phase and analyte are stable at high temperatures. Water is a highly polar eluent at low temperature and has a weak eluotropic strength resulting in a longer retention time (Al-Khateeb & Smith, 2009 Sanagi et al., 2004, Guillarme et al., 2004).

A series of eco-friendly superheated water chromatographic (SHWC) methods both with and without buffer in the mobile phase and with low percentage of organic modifiers in different elution systems have been reported for the separation of different complex species (Vidotti et al., 2006; Al-Khateeb and Smith, 2008; Smith, 2008; Al-Khateeb et al., 2021). If the stationary phase and analyte are stable at high temperatures, the separation via high-temperature liquid chromatography with thermostable stationary phases provides many advantages such as a decrease in the analysis time due to decreases in the back pressure and solvent viscosity, as well as enhancement of the mass transfer and diffusion coefficient of the analyte in the mobile phase (Al-Khateeb, 2021, Smith, 2008, Sanagi et al., 2004, Smith, 2008).

To the best of our knowledge, high-temperature chromatographic analysis of NSAIDs using a mobile phase with a low percentage of organic solvent and superheated water has not been reported till date. The present study focuses on i) achieving a short analysis time using a low percentage of organic solvent and water for reversed-phase (RP)-HPLC analysis at high temperatures, ii) validating the proposed method for the simultaneous analysis of NSAIDs [56], iii) analysing the NSAIDs in water samples and pharmaceuticals drugs, iv) determining the most probable retention mechanism using a low percentage of organic solvent and superheated water mobile phases, and v) the removal of NSAIDs from real water samples using water-rich mobile phase at high temperatures.

2

2 Experimental

2.1

2.1 Materials and methods

All solvents and chemicals were of analytical reagent (AR) grade and used as received. The pharmaceutical drugs, ibuprofen (IBU), ketoprofen (KET), naproxen (NAP), and the sodium salt of diclofenac (DIC) were standard and analytical grade (99.99% purity), and were purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade acetonitrile (ACN), methanol (MeOH) were obtained from BDH Chemicals Ltd. (Poole, UK), and HPLC-grade acetic acid and trimethylamine were purchased from Sigma-Aldrich. A stock standard solution (1 mg/mL) of each drugs was individually prepared by dissolving an appropriate weight of target compound in MeOH. More diluted standard solutions (0.1–500 mg/mL) of each drugs were also prepared in mobile phase composition.

2.2

2.2 Chromatographic conditions

The separations were carried out using an Agilent 1200 series liquid chromatography system (Agilent Technologies Inc., Santa Clara, CA, USA) equipped with a quaternary pump, autosampler, and multiwavelength detector (MWD). This device was linked to a Chemstation software that was used for data collection and analysis.

All separations were performed using an analytical RP Zir-Chrom PBD column (5 μm, 4.6 mm × 150 mm; ZirChrom Separations Inc., Anoka, MN, USA). The inlet temperature of the mobile phase was controlled by placing a preheating coil made of stainless-steel tubing (100 cm × 0.005 mm internal diameter (ID)) in the column oven before the analytical column. In addition, a stainless-steel tube was placed after the analytical column (100 cm × 0.005 mm ID) to cool the mobile phase to ambient temperature before it reached the MWD. The instrument was interfaced to a computer to obtain and analyze the data. Deionised water with a resistivity of at least 18.2 MΩ.cm was obtained using a Milli-Q Plus system (Millipore, Bedford, MA, USA). Isocratic elution was performed employing the mobile phase containing different percentages of ACN ranging from 50% to 20% with constant volumes of phosphate buffer (0.4 mL per 100 mL of solution) to control the pH. Before usage, the solvent was filtered using a 0.45-µm membrane filter and degassed ultrasonically. The sample injection volume was 10 µL, and the temperature of the column was increased to the desired temperature and equilibrated for 30 min before measurement.

2.3

2.3 Application of HTLC method in different matrices

2.3.1

2.3.1 Determination of NSAIDs in pharmaceutical formulations

Four commercially available pharmaceutical preparations of KET, NAP, DIC, and IBU in tablet forms including Ketofan (25 mg), Proxen (250 mg), Cataflam (50 mg), and Advil (200 mg), respectively, were used in this work. The stock solutions of the tablets were prepared by crushing the tablets to finely ground powders and then dissolving these in 50 mL of ultrapure water. This analysis was carried out to determine the proportion of the drugs (KET, NAP, DIC, and IBU) in these pharmaceutical drugs. All samples (pharmaceutical and spiked samples) were filtered using 0.45-µm syringe filters (Millex-GV Syringe Filter, Durapore® with a polyvinylidene fluoride membrane) fitted with disposable syringe (3 mL) to minimise the impurities in the samples before injection into the HPLC device.

The recoveries of NSAIDs from the purchased tablets were determined by comparing the claimed concentration of drugs mentioned on their boxes to the concentration found after analysis. Expressed as a percentage of the label claim, the recoveries obtained indicate that the advertised quantity of drug was present in the tablets. The recoveries ranged from 97.20% to 99.66%, having an RSD of less than 3.0%, indicating the acceptable accuracy of the method, as shown in Table 6. This good recovery indicates that there was no interference from the excipients present in the formulation.

2.3.2

2.3.2 Real water samples

Two real water samples were investigated to examine the efficacy of MWCNTs for the extraction of the four NSAIDs from real environmental water. A ground water was collected from Jazan city and a wastewater sample was obtained from the Membrane Bio-Reactor Sewage Treatment Plant (6000 m3/day, MBR 6000 STP) of King Abdulaziz University, Jeddah, Saudi Arabia (21.487954° N, 39.236748° E). The two samples were filtered through a 0.45 μm millipore membrane filter and kept in Teflon® bottles at 5 °C in the dark.

The samples were analyzed by the HTLC- RE-HPLC method for their NSAIDs content and none of the NSAIDs was detected indicating that these samples are not polluted with the aimed pharmaceuticals. Then, the ground water and wastewater treatment experiment were performed adopting the spiking technique by the addition of NSAIDs solutions to the water samples to achieve a 5.0 mg/L final concentration. Then, the pH of sample was adjusted to pH 2.0. Then, 10 mL of each sample was transferred to a glass vial, in which they were mixed with different weight ranged from 10 to 100 mg of MWCNTs for optimization the extraction experiment and left in contact for 30 min and kept at room temperature. Thereafter, the solution was filtered and analyzed by HTLC and SWLC methods for its NSAIDs content using flow rates of 1 and 2 mL min−1. For comparison, the same steps were repeated to analyze the un-purified sample by omitting the addition of MWCNTs. The % adsorption was calculated by comparison of the NSAIDs’ concentration found before and after treatment with MWCNTs.

3

3 Results and discussion

3.1

3.1 Optimisation in the method development for chromatographic separation

The most important aspects of method development in liquid chromatography include appropriate resolution of all analytes, short analysis time, and high sensitivity with low organic solvent consumption. These goals can be achieved by adjusting different chromatographic parameters to afford the desired response. The main analytical parameters that can be optimised are the stationary-phase and mobile phase compositions, pH, flow rate, and column temperature.

3.1.1

3.1.1 Effect of mobile phase composition

To examine the relationship between the proportion of organic modifier and temperature, KET, NAP, DIC salt, and IBU were analysed employing a Zir-Chrom PBD column using 50%, 40%, 30%, and 20% of acetonitrile as organic modifier in the mobile phase and 100% pure water mobile phase at different temperatures. When 50% ACN was used at a control temperature of 30 °C, the peaks for KET, NAP, and DIC salt were found to overlap at a retention time of 1.516 min (Fig. S1. Supplementary Material). With 40% MeOH mobile phase provide good resolution and long analysis time. 40% ACN, KET, NAP, DIC, and IBU peaks were resolved at 30 °C and 40 °C. However, at elevated temperature the peaks for KET and NAP overlapped at 50 °C at a retention time of 1.678 min (Fig. S2. Supplementary Material). With 30% ACN as the mobile phase, the retention times for all drugs decreased with an increase in temperature. However, the peaks for NAP and DIC did not resolve completely in the same elution order at a retention time of 1.571 min with an increase in temperature from 40 °C to 90 °C as in Fig. 1. The mobile phase with 20% ACN afforded good resolution because of decreasing in the organic solvent in the mobile phase. Retention time decrease with an increase in temperature from 40 °C to 140 °C (Fig. 1). The use of pure water also afforded good resolution at 100–140 °C for a mixture of NSAIDs as illustrate in Fig. 1.

Separation of selected NSAIDs including ketoprofen (1), naproxen (2), diclofenac sodium salt (3), and ibuprofen (4) employing mobile phases with phosphate buffer contain A) 30% ACN at 40–100 °C, B) 20% ACN-water at 40–140 °C, and C) superheated water at 100–140 °C, using a Zir-Chrom PBD column 5 μm (4.6 mm × 150 mm), Condition: flow rate of 1 mL min−1 and detection at 220 nm.
Fig. 1
Separation of selected NSAIDs including ketoprofen (1), naproxen (2), diclofenac sodium salt (3), and ibuprofen (4) employing mobile phases with phosphate buffer contain A) 30% ACN at 40–100 °C, B) 20% ACN-water at 40–140 °C, and C) superheated water at 100–140 °C, using a Zir-Chrom PBD column 5 μm (4.6 mm × 150 mm), Condition: flow rate of 1 mL min−1 and detection at 220 nm.

3.1.2

3.1.2 Effect of temperature

To determine the stability of the ZirChrom PBD column as well as the effect of temperature on the retention of different drugs, high-temperature experiments were performed by increasing the temperature from 40 °C to 140 °C in 10 °C increments using low concentrations of ACN in water and 100% water mobile phases. At low temperatures, the mobile phase is highly polar and therefore possesses a weak eluotropic strength, which results in long retention times. High temperatures can be used to reduce the back pressure, mobile phase viscosity, and dielectric constant, thereby reducing the retention on the stationary phase because of increases in the mass transfer and diffusion coefficient of the mobile phase and analyte (Cole et al., 1992, Coym & Dorsey 2004). For each mobile phase studied, a marked decrease in the retention time was observed with an increase in temperature. For example, with 20% ACN as the mobile phase, the retention times for all drugs decreased with an increase in temperature and no change in selectivity was observed in the elution order. At column temperature 40, 50, and 60 °C, DIC and IBU eluted at retention times of 7.919 and 6.856 min, respectively. However, when the temperature was increased to 70 °C, the DIC and IBU peaks resolved with retention times of 4.473 and 5.241 min, respectively. Thus, decreases in the retention times were observed from 3.26, 3.79, 7.91, and 7.91 at 40 °C to 1.47, 1.64, 1.64, and 2.13 at 140 °C for KET, NAP, DIC, and IBU, respectively (Fig. 1).

As expected, a long time was time required for the separation when a low percentage of organic solvent was used. Therefore, high temperature was employed such that the stationary phase was not adversely affected. Buffered mobile phases with pure water were investigated at high temperatures to explore and determine the efficiency of an eco-friendly separation. No overlap of peaks, good resolution, and short analysis time were observed at 140 °C (Fig. 1). The decreases in the retention times from 2.25, 2.94, 5.69, and 6.04 at 100 °C to 2.01, 2.46, 3.49, and 4.09 at 140 °C were observed for KET, NAP, DIC, and IBU, respectively (Table 1).

Table 1 Effects of different parameters on the retention and column performance in chromatographic separation using low percentages of organic solvents and superheated water.†
Mobile phases NSAIDs compounds KET NAP DIC IBU
Temperature K Rs α N k Rs α N k Rs α N k Rs α N
40 °C 0.73 1.02 1.5 3688 0.97 1.28 1.14 1374 1.71 3.21 1.76 886 2.07 0.96 1.21 834
1–30% ACN 50 °C 0.65 3.67 1.39 1733 0.82 1.16 1.1 1433 1.29 2.58 1.57 1100 1.67 1.2 1.29 1022
60 °C 0.59 4.15 1.35 2747 0.74 1.19 1.09 1380 1.08 2.12 1.45 1201 1.45 1.39 1.35 1169
70 °C 0.54 4.32 1.34 3831 0.68 4.04 1.09 1375 0.91 1.84 1.34 1249 1.28 1.86 1.4 1343
80 °C 0.48 2.67 1.27 6140 0.62 1.34 1.09 1535 0.75 0.82 1.21 1786 1.14 2.05 1.51 1631
90 °C 0.45 3.49 1.16 2338 0.57 1.16 1.08 2260 0.57 1.16 1 2260 1.01 2.31 1.76 2044
Thermodynamic parameters ΔH° (kJ/mol) ΔS° (J/mol/K) R2 ΔH° (kJ/mol) ΔS° (J/mol/K) R2 ΔH° (kJ/mol) ΔS° (J/mol/K) R2 ΔH° (kJ/mol) ΔS° (J/mol/K) R2
−9.21 –32.03 0.9973 −9.71 −31.51 0.9921 −19.64 −58.36 0.9904 −13.23 −36.49 0.9942
Temperature K Rs α N k Rs α N k Rs α N k Rs α N
40 °C 1.76 2.01 1.84 576 2.47 1.44 1.4 517 6.92 4.08 2.79 429 6.91 4.08 1 429
2–20% ACN 50 °C 1.62 1.58 1.73 537 2.23 1.37 1.37 555 5.85 3.63 2.61 312 5.85 3.63 1 312
60 °C 1.58 1.64 1.71 553 2.13 1.36 1.34 618 4.79 3.48 2.24 523 5.46 0.62 1.13 505
70 °C 1.36 2.7 1.57 891 1.78 1.37 1.3 888 3.47 4.17 1.94 835 4.24 1.39 1.22 786
80 °C 1.2 2.38 1.48 1088 1.55 1.37 1.29 1088 2.73 3.57 1.76 1070 3.52 1.71 1.28 1065
90 °C 1.02 1.85 1.41 1260 1.32 1.36 1.29 1328 2.14 2.97 1.62 1394 2.88 2.04 1.34 1376
100 °C 0.85 1.63 1.29 1329 1.1 1.41 1.29 1661 1.62 2.54 1.47 1712 2.28 2.49 1.4 1679
110 °C 0.71 1.34 1.2 1585 0.93 1.42 1.31 1886 1.29 2.18 1.38 1984 1.88 2.74 1.45 2215
120 °C 0.56 0.95 1 2908 0.83 1.44 1.48 2117 1.08 1.68 1.29 2069 1.61 3.01 1.49 2734
130 °C 0.54 3.21 0.99 1484 0.73 1.32 1.33 3901 0.88 1.31 1.21 3759 1.35 3.18 1.52 3512
140 °C 0.47 3.83 0.98 3492 0.64 1.55 1.35 2344 0.64 1.55 1 2344 1.13 3.43 1.75 3538
Thermodynamic parameters ΔH° ΔS° (J/mol/K) R2 ΔH° (kJ/mol) ΔS° R2 ΔH° (kJ/mol) ΔS° (J/mol/K) R2 ΔH° (kJ/mol) ΔS° (J/mol/K) R2
−17.19 −47.72 0.9925 −15.49 −40.87 0.9929 −25.77 −65.08 0.9923 −19.74 −46.17 0.9903
Temperature K Rs α N k Rs α N k Rs α N k Rs α N
3-Superheated water 100 °C 1.26 2.07 1.29 354 1.94 1.44 1.3 492 4.69 2.59 1.91 730 5.05 0.24 1.94 211
110 °C 1.21 1.92 1.27 464 1.88 1.54 1.29 620 4.47 4.39 1.88 921 4.47 4.39 1.88 921
120 °C 1.09 1.43 1.23 356 1.74 1.5 1.28 703 3.72 2.91 1.72 394 3.99 0.27 1.57 334
130 °C 1.02 1.76 1.21 715 1.59 1.74 1.27 904 3.05 3.55 1.56 1139 3.56 1.01 1.12 1233
140 °C 1.01 1.50 1.14 1148 1.47 0.48 1.25 1172 2.49 2.67 1.41 1563 3.09 1.45 1.16 1904
Column temperature 100 °C 110 °C 120 °C 130 °C 140 °C
Thermodynamic parameters ΔH° (kJ/mol) ΔS° (J/mol/K) ΔH° (kJ/mol) ΔS° (J/mol/K) ΔH° (kJ/mol) ΔS° (J/mol/K) ΔH° (kJ/mol) ΔS° (J/mol/K) ΔH° (kJ/mol) ΔS° (J/mol/K)
KET −8.75 –22.95 −7.59 −19.87 −6.48 −17.02 −5.43 −14.38 −4.43 −11.94
NAP −15.89 −21.59 −18.23 −27.75 −20.44 –33.45 –22.54 −38.72 −24.53 −43.62
DIC −27.68 –33.47 −34.67 −51.95 −41.29 −69.04 −47.59 −84.87 −53.59 −99.56
IBU −25.10 −37.14 −26.26 −40.22 −27.37 −43.06 −28.42 −45.70 −29.42 −48.15
† Where NSAIDs; Non-steroidal anti-inflammatory drugs; k; retention factor, N/m; column efficiency parameter; Rs; resolution between two peaks, α selectivity of column. ΔH° enthalpy of solute transferee from the mobile phase to the stationary phase and ΔS° entropy of solute transferee from the mobile phase to the stationary phase.

The results for the separation of the NSAIDs using the ZirChrom PBD column with decreasing ACN percentage (%) and increasing temperature are shown in Fig. 2. For the mobile phases with 30% ACN and 20% ACN at 60 °C and flow rate 1 mL min−1, an efficient separation of the NSAIDs with good resolution is obtained with 30% ACN owing to the solvation of the stationary phase by the organic solvent as well as temperature (Fig. 2-A). For the mobile phases containing 20% ACN at 80 °C and that containing 0% of acetonitrile which is superheated water at 140 °C, the separation of the NSAIDs occurred at comparable retention times (Fig. 2-B). However, for 20% ACN mobile phases at high temperatures, it is the effect of flow rate that improves the resolution rather than high temperature (140 °C Fig. 2-C).

Controlled separation of NSAIDs (ketoprofen (1), naproxen (2), diclofenac sodium salt (3), and ibuprofen (4)) on the 5 μm Zir-Chrom PBD column using A) Separation selected NSAIDs at 20% and 30% ACN at 60C with flow rate 1 mL/min B) Separation NSAIDs with 20% ACN at 80 °C and superheated water at 140 °C; at a flow rate of 1 mL min−1, C) Separation using 20% ACN at different flow rate.
Fig. 2
Controlled separation of NSAIDs (ketoprofen (1), naproxen (2), diclofenac sodium salt (3), and ibuprofen (4)) on the 5 μm Zir-Chrom PBD column using A) Separation selected NSAIDs at 20% and 30% ACN at 60C with flow rate 1 mL/min B) Separation NSAIDs with 20% ACN at 80 °C and superheated water at 140 °C; at a flow rate of 1 mL min−1, C) Separation using 20% ACN at different flow rate.

This effect is attributed to the mass transfer at high temperature (Fig. 2) and indicates that a high temperature and flow rate can replace the use of high percentage of organic component in the mobile phase. Thus, the retention can be controlled either by varying the percentage of the organic solvent in the mobile phase or by using high column temperature and flow rate [53].

As shown in Fig. 1-B, the column temperature is an important parameter for the separation of selected compounds. At high column temperatures, the resolution decreases, peaks become narrow, analysis time decreases, and peak height increases. At a column temperature of 120 °C, sufficient resolution is obtained and a significant improvement in the peak shape of IBU is observed with 20% ACN. Thus, 20% ACN at a temperature of 120 °C was selected as the solvent and column temperature for optimising the separation and analysis of NSAIDs.

Table 1 shows that the retention factor decreases with an increase in temperature. In addition, no change in the order of the elution of the selected compounds is observed with all mobile phases investigated in this study. As expected, the retention time increases when a low percentage of organic modifier is used (Fig. 2) while maintaining the same elution order because of the poor solvation of stationary phase. The analyte peaks broaden with a decrease in the percentage of the organic solvent in the mobile phase, with an increase in the analysis time. However, the resolution of the compounds increases with a decrease in the ACN percentage in the mobile phase and decreases with an increase in the column temperature.

As shown in Table 1, the column efficiency for the separation of selected anti-inflammatory drugs improves with an increase in the column temperature, particularly for the compounds that elute towards the end (DIC and IBU). Furthermore, the theoretical number of plates increases, and the peak shape improves with an increase in temperature until the Zir-Chrom PBD column limit is reached at 140 °C. The improved efficiency is due to a decrease in the mobile phase viscosity at elevated temperatures, which increases the diffusivity of the mobile phase and analyte, as diffusivity is inversely proportional to viscosity (Guillarme et al., 2004b). Thus, narrow bands and high column efficiency are observed. For the compounds eluting early (KET and NAP), an increase in temperature results in a decrease in the column efficiency. This is probably because of the interaction of the poorly retained compounds with the wall of the separation column; additionally, an increase in the temperature during separation results in a low mass transfer resistance factor and high longitudinal diffusion, which become the dominant factors (Heinisch & Rocca, 2009).

The peak width and height are also affected by an increase in the column temperature. Upon increasing the column temperature, the peak height increases and the peak width and tailing decrease, particularly for the compounds that elute early, as shown in Table 1 and Fig. 1. The effect of temperature on selectivity when low percentages of ACN are used in the mobile phase is shown. The selectivity decreases with an increase in temperature and increases with a decrease in the percentage of ACN in the mobile phase (Li & Carr, 1997). The peak resolution (Rs) values of various compounds decrease with an increase in temperature and increase with an increase in the percentage of the organic component in the mobile phase. As shown in Table 1, the resolution values of KET and NAP at 40 °C increase from 1.02 and 1.28 using 30% ACN, to 2.01 and 1.44, respectively, with 20% ACN at 100 °C; correspondingly, the resolution values increase from 1.63 and 1.41 to 2.07 and 1.44, respectively, with pure water.

Retention mechanism

To understand the effect of temperature on the retention behaviour of each compound, the thermodynamic behaviour was investigated using the van’t Hoff relationship (ln k vs. 1/T), as shown in Fig. 3, with superheated water as the mobile phase. For mobile phase compositions with 30% and 20% ACN, the van’t Hoff plots are linear with a high correlation coefficient (r2) of > 0.99. The enthalpy (ΔH°) for the solute transfer from the mobile to the stationary phase can be calculated from the slope, and the entropy (ΔS°) can be obtained from the intercept of the data for each compound. As no significant deviation from linearity is observed in the van’t Hoff plots in the temperature range investigated, it can be assumed that there is no change in the retention mechanism of the studied compounds when 30% and 20% ACN are used as mobile phases with the Zir-Chrom PBD column. For 30% ACN, the enthalpy values are −1104.71, −1158.97, −2340.02, −1582.47 kJ mol−1 for KET, NAP, DIC, and IBU, respectively. For 20% ACN, the corresponding enthalpy values are −2052.99, −1850.57, −3075.83, and −2350.68, respectively. When 100% water is used as the mobile phase, the van’t Hoff plot is nonlinear, and the enthalpy values for KET, NAP, DIC, and IBU at 100 °C are 4140.02, −12845, −35304, and −8372.2 kJ mol−1, respectively. At 140 °C, the corresponding enthalpy values are 4140. 02, −12848, −35305 and −8372.3, respectively (Table 1). As expected, the ΔH° values become more negative with a decrease in the ACN content in the mobile phase. This indicates a strong retentive interaction between the mobile and stationary phases when the percentage of the organic component in the eluent is low, and the hydrophobic interactions dominate the separation. This is consistent with the observations reported in prior studies (Li & Carr, 1997, Al-Khateeb, 2019). In contrast, the entropy values are −3.842, −3.75966, −6.95172, and −4.3883 J mol−1 K−1 for KET, NAP, DIC, and IBU, respectively, with 30% ACN, and with 20% ACN, the corresponding entropy values are −5.69626, −4.8812, −7.76723, and −5.49874 J mol−1 K−1, respectively. Using 100% H2O, the entropy values at 100 °C were 5.7617, 5.97072, 5.96753, and 5.99408 J mol−1 K−1 for KET, NAP, DIC, and IBU, respectively, while at 140° C, these values are 5.7618, 5.9707, 5.97043, and 5.99407 J mol−1 K−1, respectively. Thus, for all mobile phase compositions, both ΔH° and ΔS° values are negative for all solutes under the experimental conditions, and the retention is exothermic. A significant increase in enthalpy is observed at high temperatures, which is greater than the entropy; this indicates that the retention mechanism is enthalpically driven at high mobile phase temperatures (Flieger et al., 2019).

Vanʹt Hoff plots of NSAIDs using water-rich solvent 30%ACN (A), 20% ACN(B) and superheated water mobile phase (C).
Fig. 3
Vanʹt Hoff plots of NSAIDs using water-rich solvent 30%ACN (A), 20% ACN(B) and superheated water mobile phase (C).

The magnitudes of the interactions of the methyl and carbonyl groups with the stationary phase of each solute differ between various NSAIDs. This indicates that the enthalpy increases through strong interactions with the stationary phase (Guillarme et al., 2004, Heinisch & Rocca, 2009, Karadurmus et al., 2019). Accordingly, the enthalpy for IBU increases owing to the strong interactions of the methyl groups with the stationary phase. However, when 100% water is used, the van’t Hoff plot for the separation of NSAIDs exhibits a nonlinear relationship, which indicates that the enthalpy and entropy are temperature dependent, as shown in Fig. 3-C. The negative enthalpy values indicate that it is energetically favourable for the solute to remain in the stationary phase and the positive enthalpy value for KET indicates that it is energetically less favourable for the solute to remain in in the stationary phase. In addition, the positive values of entropy decrease in the elution order observed in the chromatogram as the solute transfer occurs from the mobile to the stationary phase.

3.1.3

3.1.3 Effect of flow rate

For isocratic separation, the flow rate and retention time are generally inversely correlated; as the flow rates increases, the retention decreases, as shown in Fig. 4. The optimum flow rate is dependent on the particle size of the column packing material, which affects the efficiency of analyte separation. An increase in the flow rate increases the back pressure of the system, which limits the pump performance. High temperature reduces the back pressure and allows the separation at high flow rates using stationary phases with small particle sizes (Heinisch et al., 2009, Al-Khateeb, 2019).

Van Deemter relationship for NSAIDs Separation conditions include 20% ACN with column temperature 120 °C at 220 nm detection wavelength and flow rate of 1 mL min−1.
Fig. 4
Van Deemter relationship for NSAIDs Separation conditions include 20% ACN with column temperature 120 °C at 220 nm detection wavelength and flow rate of 1 mL min−1.

In this study, the effect of an increase in the flow rate of the mobile phase from 0.5 to 2.8 mL min−1 at optimised conditions (20% ACN and 120 °C) on the Zir-Chrom PBD C18 column was investigated. With an increase in the flow rate, the analyte retention decreases. In addition, the resolution between the two adjusted compounds keto and napo decreases with an increase in flow rate. A considerable loss of efficiency is observed when a high flow rate is used, although the peaks are resolved. Thus, the height equivalent to a theoretical plate increases in the van Deemter curve. The optimum flow rate is 1.0 mL min−1, and at a higher flow rate than this value, the efficiency decreases, as shown in Fig. 4. Based on the curve shape, the effect of longitudinal diffusion (B-term in the van Demeter equation) appears to be dominant at low linear velocities as the flow rate increases beyond the optimum value.

3.2

3.2 HTLC method validation

To evaluate the reliability of the established high temperature liquid chromatography (HTLC) method at high water rich and pure water for analysis of the four investigated drugs, different parameters including linearity, range, limit of quantification (LOQ), limit of detection (LOD), accuracy, precision, sensitivity, and recovery were thoroughly investigated as guided by USP 26 and ICH Q2R1 guideline (United States Pharmacopeial Convention, 2003, CPMP/ICH/381/95, 1994, Ozgur, 2013, Sendanyoye, 2018).

Linear relations were accomplished by graphing the mean peak areas against drug concentration over the ranges of 500.0–0.1, 500.0–0.005, 500.0–0.01 and 500.0–0.1 mg/L for KET, NAP, DIC, and IBU, respectively, with correlation coefficient r ≥ 0.996. The LODs and LOQs were also calculated as 3.3σ/a and 10σ/a, respectively, where (σ) is the standard deviation (SD) of the response and (a) is the slope of the regression line. (Miller and Miller, 2000) A summary of the obtained results is presented in Table 2.

Table 2 Figures of merits of the developed method for analysis of KET, NAP, DIC, and IBU.†
NSAIDs †† compound KET NAP DIC IBU
Linearity (mg/L) 0.1–500 0.005–500 0.01–500 0.01–500
R2 0.993 0.996 0.999 0.999
Slope 21.753 76.062 31.256 22.686
Intercept 84.083 149.27 52.029 40.431
LOD (µg / L) 14.0 2.0 4.2 32.0
LOQ (µg/ L) 44.8 8.0 12.0 98.0
† Mobile phase composed of 20% acetonitrile with phosphate buffer at 120 °C employing Zirconia-C18 column (5 µm; 150 mm × 4.6 mm ID).
†† Ketoprofen = KET; Naproxen = NAP; Diclofenac salt = DIC; Ibuprofen = IBU.

Inter-day and intra-day variations were used to test the precision of the proposed procedures for the analysis of four drugs at the 10 mg/L concentration in a single day and on 3 successive days, respectively, the results are summarized in Table 3. The intra-day repeatability values show 0.08–1.52% RSD, whereas the inter-day precision values show 1.01–2.67% RSD. The % RSD values were ≤ 2.7 for the four compounds (Table 3).

Table 3 Inter-day and intra-day precision of NSAIDs drugs (10 mg/L) by the developed method.††
NSAIDs†† drugs Accuracy (%) Intra-day precision (n = 3) Inter-day precision (n = 9)
Mean %RSD Mean %RSD
Day1 Day2 Day3 Day1 Day2 Day3
Ketoprofen 97.8 506.83 508.37 505.84 1.52 0.22 1.25 505.37 1.01
Naproxen 99.6 2225.29 2230.06 2227.12 0.11 0.08 0.14 2203.72 1.37
Diclofenac salt 98.1 790.27 789.69 790.48 0.09 0.70 0.71 801.43 2.67
Ibuprofen 102.8 427.30 428.38 428.02 0.12 0.32 0.14 432.54 1.72
†† Mobile phase composed of 20% acetonitrile with phosphate buffer at column temperature 120 °C employing Zirconia-C18 column (3.5 µm; 150 mm × 4.6 mm ID).

Moreover, to validate the reliability of HTLC method for the determination of the investigated analytes in water samples from distinct sources by analyzing known concentrations (2.0–100.0 mg/L) of the four drugs spiked into the water samples. The experimental results show that the concentrations of the four compounds are below the LOD of the developed method. Therefore, spiking experiments were performed using different concentrations of the four compounds. The recoveries of different known added amounts of the analytes to wastewater and ground water samples were calculated. As deduced from Table 4, the good % recoveries of the four compounds ranged from 96.2 to 100.0% with %RSD values are 1.1–3.6% for the ground water and 1.6–4.3% for wastewater, indicating acceptable performance of the developed method (Table 4). The amounts determined by the proposed method are in agreement with those obtained by the standard methods (Baranowska, 2012) reported for nonsteroidal components. The experimental results for the student t (texp.: 1.72–2.27) and F (Fexp: 1.2–2.33) tests at 95% confidence (n = 5) do not exceed the tabulated t (2.31) and F (6.38) values, indicating the precision of the method.

Table 4 Recovery of the four NSAIDs form different-types of water samples by the developed method.††
NSAIDs
drug†† 		Spiked
Concentration (mg/L) 		Ground water
Recovery%±SD (n = 3) 		Wastewater
Recovery%±SD (n = 3)
KET 5.0 97.2 ± 3.6 97.1 ± 4.3
10.0 98.6 ± 2.3 96.4 ± 2.8
50.0 98.4 ± 1.7 97.6 ± 1.9
NAP 2.0 97.9 ± 2.9 96.02 ± 3.9
10.0 99.1 ± 2.1 96.2 ± 2.8
50.0 97.5 ± 1.1 96.4 ± 2.2
DIC 2.0 97.4 ± 2.5 96.5 ± 3.6
10.0 97.8 ± 2.2 96.3 ± 2.5
100.0 100.1 ± 1.4 99.3 ± 2.1
IBU 5.0 95.5 ± 2.8 96.2 ± 3.2
10.0 97.6 ± 1.3 98.8 ± 1.7
100.0 100.3 ± 1.5 99.9 ± 1.6
†† Ketoprofen = KET; Naproxen = NAP; Diclofenac salt = DIC; Ibuprofen = IBU.

Further, the recoveries of NSAIDs from the purchased tablets (Brufen, Profenid, Proxen, and Voltaren Retard) were determined by comparing the claimed concentrations of the drugs mentioned on the packaging to the concentrations measured upon analysis. Expressed as a percentage on the label, the recoveries obtained indicate that the claimed quantities of the drugs are present in the tablets. The recoveries range from 99.2 to 100.08% with an RSD of <3.0%, indicating the acceptable accuracy of the method, as shown in Table 5 (single drug tablets) and Table 6 (combination drugs). The good recovery results indicate that there is no interference from the excipients present in the formulation for 20% ACN at 120 °C (Fig. 5) and superheated water at 140 °C witflow rate 1 mL/min (Fig. 6).

Table 5 Recovery data for medicine single tablets using developed method.††
Mobile phase Buffer 20% ACN at 120 °C
Drugs †NSAID compounds Claimed (mg) Measured (mg) %Recovery %RSD
Ketofan KET 25 25.01 99.96 1.09
Proxen NAP 250 250 99.99 0.75
Cataflam DIC salt 50 50 99.99 0.22
Advil IBU 200 200.01 99.99 0.15
Mobile phase Buffer 100% H2O at 130 °C
Drugs †NSAID compounds Claimed (mg) Measured (mg) %Recovery %RSD
Ketofan KET 25 25.02 99.92 0.21
Proxen NAT 250 250.02 99.99 1.49
Cataflam DIC salt 50 50 99.99 0.52
Advil IBU 200 200.03 99.98 1.08
†† Drugs were purchased from the local pharmacy of Jeddah, Saudi Arabia. The analysis was performed using the developed method by employing Zirconia-C18 column (5 µm; 150 mm × 4.6 mm ID) with a flow rate of 1 mL min−1 and detection at 220 nm.
† NSAIDs Compound: KET: Ketoprofen; NAP: Naproxen; DIC: Diclofenac sodium salt; IBU: Ibuprofen.
Table 6 Recovery data for the combination drug tablets obtained using the developed method.††
Mobile phase 20% ACN at 120 °C
Drugs NSAID compounds Claimed (mg) Measured (mg) %Recovery %RSD
KET DIC KET DIC KET DIC KET DIC
Ketofan, Cataflam KET, DIC 25 50 24.97 50.01 100.08 99.97 0.25 0.15
NAP IBU NAP IBU NAP IBU NAP IBU
Proxen, Advil NAP, IBU 250 200 250.22 200.02 99.9 99.98 0.06 2.46
Mobile phase 100% H2O at 130 °C
Drugs NSAID compounds Claimed (mg) Measured (mg) Recovery% %RSD
KET DIC KET DIC KET DIC KET DIC
Ketofan, Cataflam KET, DIC 25 50 25.02 50.01 99.88 99.97 0.07 0.69
NAP IBU NAP IBU NAP IBU NAP IBU
Proxen, Advil NAP, IBU 250 200 200.04 250.02 99.98 99.97 1.06 2.41
†† Drugs were purchased from the local pharmacy of Jeddah, Saudi Arabia. Analysis was performed using the developed method by employing the Zirconia-C18 column (5 µm, 150 mm × 4.6 mm ID), Buffer mobile phase contain with phosphate buffer with a flow rate 1 mL min−1 and detection at 220 nm. NSAIDs Compound: KET: Ketoprofen; NAP: Naproxen; DIC: Diclofenac sodium salt; IBU: Ibuprofen.
HPLC chromatograms of the standard NSAIDs and tablet of single drug and combination drugs in tablet employing 20% ACN at 120 °C on Zir-Chrom PBD 5 μm column at a flow rate of 1 mL min−1 and MWD detection at 220 nm.
Fig. 5
HPLC chromatograms of the standard NSAIDs and tablet of single drug and combination drugs in tablet employing 20% ACN at 120 °C on Zir-Chrom PBD 5 μm column at a flow rate of 1 mL min−1 and MWD detection at 220 nm.
HPLC chromatograms of the standard NSAIDs and tablet of single drug and combination drugs employing superheated water at 140 °C as mobile phases on Zir-Chrom PBD column at a flow rate of 1 mL min−1 and MWD detection at 220 nm.
Fig. 6
HPLC chromatograms of the standard NSAIDs and tablet of single drug and combination drugs employing superheated water at 140 °C as mobile phases on Zir-Chrom PBD column at a flow rate of 1 mL min−1 and MWD detection at 220 nm.

3.2.1

3.2.1 Application for purification of water samples

Application of the developed method for water purification was demonstrated by evaluation on two real samples include ground water and a wastewater sample collected from our institutional treatment plant. Experimental results showed that none of the four compounds was detected. Therefore, spiking experiments were performed at a final concentration of 5.0 mg/L of the four NSAIDs compounds. By comparison of the found concentration before and after treatment with MWCNTs, the % adsorption of the targeted compounds are illustrated in Fig. S3. The percentage extraction by the MWCNTs was investigated using 5.0 mg L−1 of KET, NAP, DIC, and IBU, and the results obtained for the optimisation of the removal by the CNTs are shown in Fig. 3S. As shown, the percentage extraction increases significantly for KET, NAP, DIC, and IBU with an increase in the MWCNT mass until a maximum is reached at 100 mg of MWCNTs. The percentage removal increases from 33.4, 39.6, 66.3, and 28.4% when 10 mg of MWCNTs are used, to 81.3, 92.2, 97, and 60.8%, with 100 mg of MWCNTs for KET, NAP, DIC, and IBU, respectively. This enhancement in the percentage removal with an increase in the MWCNT mass is mainly due to the availability of additional binding sites on the MWCNT surfaces for the removal and adsorption of the NSAID compounds.

To determine the practical applicability of this method, it was employed for the analysis of real wastewater and ground water samples as well as some well-known commercial drugs of the studied NSAIDs. The accuracy was determined by obtaining three replicate measurements at three spiked concentration levels in the real samples. The recoveries range from 90.48% to 98.15% for the wastewater and from 93.50% to 97.77% for ground water, as shown in Fig. 7.

Removal of NSAIDs from ground and wastewaters employing A: 20% ACN at 120 °C and B: superheated water at 130 °C with different flow rates of 1 mL min−1 and 2 mL min−1.
Fig. 7
Removal of NSAIDs from ground and wastewaters employing A: 20% ACN at 120 °C and B: superheated water at 130 °C with different flow rates of 1 mL min−1 and 2 mL min−1.
Removal of NSAIDs from ground and wastewaters employing A: 20% ACN at 120 °C and B: superheated water at 130 °C with different flow rates of 1 mL min−1 and 2 mL min−1.
Fig. 7
Removal of NSAIDs from ground and wastewaters employing A: 20% ACN at 120 °C and B: superheated water at 130 °C with different flow rates of 1 mL min−1 and 2 mL min−1.

3.2.2

3.2.2 Evaluation of superheated water chromatography procedure greenness

The twelve principles of green analytical chemistry were appraised using the analytical greenness software: AGREE which was developed recently (Pena-Pereira et al., 2020). This metric considers sample procedure, sample size, device position, stages of preparation automation, miniaturization, derivatization, generation and handling of waste, consumption of energy, source and toxicity of reagents, and operator’s safety. The software powers a scoring system that converts the principles into numerical values and gives a final score within the range of 0 to 1, where 1 is assigned to the ideally green method that met all the criteria. In the current study, application of this software to evaluate the developed technique showed excellent greenness with a score of 0.96 as show in Fig. 8.

Generic result of assessment (left) and the corresponding color scale for reference (right).
Fig. 8
Generic result of assessment (left) and the corresponding color scale for reference (right).

As distinguished from Table 7 and the proposed method possesses the highest analytical eco- scale score among the reported literature for analysis NSAIDs.

Table 7 Comparison of the performance of the developed method and reported literature for analysis and determination NSAIDs.
Analytical technique for NSANDs RP-HPLC chromatographic condition Mobile phase Retention time Remarkable References
High temperature-RPHPLC with three 2 µm C18 columns
For KET, DIC, IBU		 Three Zorbax SB C18 at columns temperature 80 °C gradient elution ACN:H2O 0.5% acetic acid, with flow rate 1.1 mL/min KET:21.9 min, DIC:25.6 min, IBU:27.9 min Solid phase extraction flow three Zorbax SB C18 columns and gradient eluation Shaaban et al., 2011
RP-HPLC diclofenac sodium in tablet symmetry C18 column (4.6 mm × 150 mm, 3 μm) at room temperature 35% of 0.05 M orthophosporic (pH 2.0) and 65% ACN 2 min at 2 mL min 1 Costly (HPLC-solvent) Use of high percentage of hazard solvent Alquadeib, 2019
HPLC- UV
KET,NAP, DIC,IBU		 Zorbax Eclipse XDB-C18, 5 µm
LOD: 6–74 µgL− 		Isocratic:6.9 mmol/L acetic acid : 35% CAN
Gradient: A: CAN, B: ACN/ 6.9 mmol/L acetic acid		 Isocratic retention time ::55 min, and Gradient retention 32 min Water analysis by extraction using polymeric Strata X cartridge Stafiej, et al., 2007
SPE- HPLC
KET,IBU		 Kinetex Evo C18
0.8 mL/min
0.003 and 0.01 LOD:µg/mL		 50 %ACN:50%
H2O with phosphate buffer		 14 min. Need extraction costly HPLC solvent Milanetti et al., 2019
HPLC-UV
NAP, DIC,IBU		 Lichrospher C18, 5 μm
Linear detection range: 1–10 mg/L
LOD:0.04–0.13 µg/mL		 60% acetonitrile and 40% 0.2% formic acid in water 6 min at 0.8 mL min 1 Need organic solvent for extraction detection wavelength program is needed Madikizela et al., 2017
HPLC-UV
NAP, DIC,IBU		 ODS Zirconia RP-C18, 5 μmLinear detection range: 0.01–500 mg/L Buffered superheated water chromatography 5 min at 1 mL min 1 High temperature used to improved mass transfer and analyte diffusion with green separation This method

4

4 Conclusions

Herein, a simple, rapid, efficient, reliable, and sensitive RP-HPLC method is developed and validated for the analysis of NSAIDs at elevated temperatures. Superheated water-Zirconia-PBD chromatography system is effectively utilised for the rapid separation and removal of anti-inflammatory 2-arylpropionic acid derivatives using water as a green and economic eluent. Statistical analysis reveals that the high-temperature method is suitable for the analysis of NSAIDs in water samples and pharmaceutical formulations without any interference from the excipients.

An SHWC approach was developed and validated for the efficient separation of the four NSAIDs compounds. The approach was applied for the determination of these analytes in environmental, and pharmaceutical samples with excellent recoveries. The greenness of the developed approach was also positively assessed using the AGREE software. The proposed method is an excellent addition to the pre-existing analytical methods for the analyses of such compounds in terms of its effectiveness and speed, greenness and suitability of the ٍأSHWC method for diverse applications for the determination of these groups of frequently prescribed NSAIDs.

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Appendix A

Supplementary material

Supplementary data to this article can be found online at https://doi.org/10.1016/j.arabjc.2021.103226.

Appendix A

Supplementary material

The following are the Supplementary data to this article:

Supplementary data 1


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